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Pessoa envisions an approach to neuroscience that treats the brain as an "interactionally complex system": a system that cannot be understood through analysis and manipulation of its parts. I provide reason to support Pessoa's overall approach while putting pressure on some of the specific claims., (© 2022 Massachusetts Institute of Technology.)

Rationale: Increased cytokine response is common in patients receiving extracorporeal life support and is often a poor prognostic indicator. There is interest in using adjunctive cytokine adsorption technologies to reduce inflammatory burden, However, it is debated whether extracorporeal membrane oxygenation (ECMO) itself provides therapeutic benefit beyond gas exchange. Thus, we sought to characterize the inflammatory profile of ECMO in the first 72-96 h of and quantify its effect on cytokine levels in a case series of patients undergoing ECMO., Methods: Eight patients initiating ECMO were studied. Of these, we measured cytokines pre- and post-oxygenator over 96 h. Comparisons of cytokine levels were made across the oxygenator and over time., Results: The average age of patients was 64.3 years with 62% being male. Centrally cannulated patients had higher IL-6 levels (820.43 vs. 6907.53 pg/ml, p < 0.03), whereas peripherally cannulated patients had higher IL-12p70 levels (7.73 vs. 2.59 pg/ml, p < 0.05). Cytokine levels on day one included IL-12p70 (4.17 ± 2.56), IL-6 (4971.23 ± 8569.88), TNF (undetected), IL-8 (346.68 ± 670.18), IL-1B (undetected), and IL-10 (72.27 ± 87.9). Cytokine levels increased over 96 h; however, no significant differences were appreciated despite blood product transfusion. On day 3, IL-12p70 levels were significantly lower post-oxygenator (p < 0.05)., Conclusion: The inflammatory profile of ECMO does not change significantly over the early course of illness when accounting for transfusion. However, the decrease in IL-12p70 specifically at day 3 of ECMO may indicate adsorption of specific inflammatory markers by the oxygenator although the clinical significance of this is still unknown. Further investigation of the oxygenator on cytokine response is warranted., (© 2021 International Center for Artificial Organs and Transplantation and Wiley Periodicals LLC.)

Background: The purpose of this study was to assess the occupational SARS-CoV-2 infection risk among health care workers (HCW) at University of Kentucky HealthCare (UKHC) by evaluating the prevalence of SARS-CoV-2 antibodies., Methods: This is a prospective cohort study of HCW at UKHC. SARS-CoV-2 IgG antibody seropositivity was measured in a CLIA-certified laboratory utilizing the Abbott Architect SARS-CoV-2 IgG antibody assay. Demographics and work type were self-reported by study participants via an emailed survey., Results: The overall antibody positivity rate of HCW was 1.55% (5/322; 95% confidence interval: 0.65%-3.71%) at cohort entry. There were no differences in antibody positivity between those that worked directly with SARS-CoV-2 infected patients and those that did not. The antibody rate of positivity of patients during the same time period was similar, 1.8% (9/499; 95% confidence interval 0.94%-3.45%)., Conclusions: Antibody positivity was low and similar between HCW and patients tested during a similar time period. HCW positivity rates did not appear to be impacted by caring for known SARS-CoV-2 infected patients suggesting that appropriate use of personal protective equipment is effective in protecting individuals from transmission., (Copyright © 2021 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Background: For women undergoing median sternotomy, especially those with a bra cup size C or larger, breast support can reduce pain, wound breakdown, and infection. This study addressed a gap in research, identifying the best bra after sternotomy in terms of patient satisfaction and wear compliance., Objectives: To evaluate larger-breasted women's satisfaction and compliance with wearing 3 commercially available front-closure bras-with a hook-loop closure (the hospital's standard of care), a zipper closure, or a hook-eye closure-after cardiac surgery., Methods: This study used a posttest-only, 3-group randomized controlled design. A convenience sample of participants were sized and randomly assigned a product that was placed immediately postoperatively. Participants agreed to wear the bra at least 20 h/d until the provider cleared them for less wear. At inpatient day 5 or discharge, and at the follow-up outpatient visit, subjects completed investigator-developed surveys. Data were analyzed from 60 participants by using the χ2 test and Kruskal-Wallis analysis of variance; also, patterns were identified within written comments., Results: Participants were most satisfied with the hook-eye front-closure product before (P = .05) and after (P = .02) discharge. Participants recommended the hook-eye and zipper products over the hook-loop bra (H = 8.39, P = .02). Wear compliance was strongest in the group wearing the hook-eye bra., Conclusions: The hook-eye closure product had the most satisfaction and greatest wear compliance, and it received the highest recommendation. A practice change was made to fit and place the hook-eye bra in the operating room immediately after surgery., (©2021 American Association of Critical-Care Nurses.)

Objectives: This qualitative study was designed to describe the experiences of mentors for a nursing research fellowship at a southeastern US academic medical center., Background: Mentoring is an important aspect of nursing and is a key strategy to develop nurse leaders and faculty. Research mentors have been identified as essential for novice researchers to be able to complete clinical studies. However, there has been limited research on the experience of nursing research mentors, despite the emphasis on the critical nature of this role., Methods: Eight mentors for 23 studies over 8 years of a nursing research fellowship participated in a study outlining their experiences as research mentors., Results: Common themes emerged from mentors' identification of key components in the research mentoring process and elements necessary for effective support of frontline nurse researchers., Conclusions: Recommendations for developing effective, confident research mentors are drawn from the analyses.

Allogeneic hematopoietic stem cell transplant (allo-HSCT) is often used to treat acute leukemia or defects of hematopoiesis. Its widespread use is hampered by graft-vs.-host disease (GVHD), which has high morbidity and mortality in both acute and chronic subtypes. Chronic GVHD (cGVHD) occurs most frequently in skin and often is characterized by pathogenic fibrosis. Mast cells (MCs) are known to be involved in the pathogenesis of other fibrotic diseases. In a murine model of cGVHD after allo-HSCT, C57BL/6J recipients of allogeneic LP/J donor cells develop sclerodermatous dermal cGVHD which is significantly decreased in mast cell-deficient B6.Cg-Kit W-sh/ HNihrJaeBsmGlliJ recipients. The presence of MCs is associated with fibrosis, chemokine production, and recruitment of GVHD effector cells to the skin. Chemokine production by MCs is blocked by drugs used to treat cGVHD. The importance of MCs in skin cGVHD is mirrored by increased MCs in the skin of patients with dermal cGVHD. We show for the first time a role for MCs in skin cGVHD that may be targetable for preventive and therapeutic intervention in this disease., (Copyright © 2019 Strattan, Palaniyandi, Kumari, Du, Hakim, Huang, Kesler, Jennings, Sturgill and Hildebrandt.)

Background: Experimental and preclinical evidence suggest that adoptive transfer of regulatory T (Treg) cells could be an appropriate therapeutic strategy to induce tolerance and improve graft survival in transplanted patients. The University of Kentucky Transplant Service Line is developing a novel phase I/II clinical trial with ex vivo expanded autologous Treg cells as an adoptive cellular therapy in renal transplant recipients who are using everolimus (EVR)-based immunosuppressive regimen., Methods: The aim of this study was to determine the mechanisms of action and efficacy of EVR for the development of functionally competent Treg cell-based adoptive immunotherapy in transplantation to integrate a common EVR-based regimen in vivo (in the patient) and ex vivo (in the expansion of autologous Treg cells). CD25 Treg cells were selected from leukapheresis product with a GMP-compliant cell separation system and placed in 5-day (short) or 21-day (long) culture with EVR or rapamycin (RAPA). Multi-parametric flow cytometry analyses were used to monitor the expansion rates, phenotype, autophagic flux, and suppressor function of the cells. phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway profiles of treated cells were analyzed by Western blot and cell bioenergetic parameters by extracellular flux analysis., Results: EVR-treated cells showed temporary slower growth, lower metabolic rates, and reduced phosphorylation of protein kinase B compared with RAPA-treated cells. In spite of these differences, the expansion rates, phenotype, and suppressor function of long-term Treg cells in culture with EVR were similar to those with RAPA., Conclusions: Our results support the feasibility of EVR to expand functionally competent Treg cells for their clinical use.

Singh's cultural evolutionary theory posits that methods of inducing shamanic altered states of consciousness differ, resulting in profoundly different cognitive states. We argue that, despite different methods of induction, altered states of consciousness share neurophysiological features and cause shared cognitive and behavioral effects. This common foundation enables further cross-cultural comparison of shamanic activities that is currently left out of Singh's theory.

This report presents a falsely incompatible B cell crossmatch by flow cytometry in a lung transplant recipient. The patient was a 35-year-old Caucasian male with end-stage lung disease secondary to cystic fibrosis whose pretransplantation serologic workup did not disclose the presence of anti-HLA class II antibodies by single antigen bead testing. Unexpectedly, crossmatch of recipient sera with pronase-treated donor lymphocytes resulted in antibody binding to B cells only. The positive reactivity was reproducible in pronase-treated autologous B cells. Recipient sera did not react with nontreated donor or autologous lymphocytes. Herein, we describe our approach to this unexpected crossmatch result and consider the implications of false-positive crossmatch results on transplantation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Purpose: To evaluate the long-term results of proximal interphalangeal (PIP) joint surface replacement arthroplasty for arthritis using the SR PIP implant (Small Bone Innovations, New York, NY)., Methods: This is a long-term retrospective analysis of results in 39 of 43 joints first reported in 2008. Subjective results were based upon a mailed questionnaire. Active range of motion was measured by a certified hand therapist, and x-rays were obtained to analyze changes occurring since the first study., Results: The average follow-up time was 9.3 years. The average active PIP joint arc of motion in the present cohort of patients went from 64° at the first report (2008) to 56° at this time. Radiographic comparisons revealed no major changes since the first study. Ten of 11 revisions were done for pain due to loosening and were performed at an average of 20 months after the primary procedure. No further revisions were necessary in the interim. Overall, subjective measures of satisfaction and symptomatic and functional improvement remained unchanged., Conclusions: Surface replacement arthroplasty using the SR PIP implant continues to be an option for patients with osteoarthritis of the PIP joint. Long-term subjective and objective outcomes are comparable to those reported using other implants. This and other studies suggest that this procedure is not appropriate for most rheumatoid joints. In the interim between studies, we saw a reduction in the average PIP joint arc of motion, although this change did not reach statistical significance. Our original revision incidence of 26% has not changed. Subjective evaluation and radiologic findings did not change between studies., Type of Study/level of Evidence: Therapeutic IV., (Copyright © 2015 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.)

Purpose: Activating germline mutations in CARD11 have recently been linked to a rare genetic disorder associated with congenital B cell lymphocytosis. We describe a patient with a similar clinical phenotype who had a de novo germline G123D CARD11 mutation., Methods: Whole exome sequencing was performed on DNA from the patient and his biological parents. Laboratory studies examined characteristics of the patient's B and T lymphocytes. A CARD11 cDNA containing the mutation was transfected into a lymphocyte cell line to gain an understanding of its function. RNA sequencing was performed on samples from the patient and from patients with alternate germline CARD11 mutations and differential gene expression analysis was performed., Results: The patient had a decade-long history of severe polyclonal B lymphocytosis in the 20,000-90,000 lymphocytes/mm(3) range, which was markedly exacerbated by EBV infection and splenectomy at different times. He had a heterozygous germline CARD11 mutation causing a G123D amino acid substitution, which was demonstrated to induce NF-κB activation in unstimulated lymphocytes. In contrast to previous patients with CARD11 mutations, this patient's B cells exhibited higher expression of several cell cycle progression genes, as well as enhanced proliferation and improved survival following B cell receptor stimulation., Conclusions: This is the third reported germline and first de novo CARD11 mutation shown to cause congenital B cell lymphocytosis. The mutation was associated with a dramatically greater lymphocytosis than in previously described cases, disproportionate to the level of constitutive NF-κB activation. However, comparative review of the patient's clinical history, combined with additional genomic and functional analyses, underscore other important variables that may affect pathophysiology or regulate mutant CARD11 function in B cell proliferation and disease. We now refer to these patients as having BENTA disease (B cell Expansion with NF-κB and T cell Anergy).

Background: Alloantibodies to the Bw4 epitope are known to be heterogeneous, but it is widely assumed that anti-Bw4 alloantibodies arise only in individuals who do not express a Bw4 epitope., Methods: Bw4 expression was confirmed by DNA sequence analysis. Anti-Bw4 reactivity was confirmed by absorption with transfected cells., Results: A Bw4 (B*27:05 or B*27:13) patient expressed antibody that bound all Bw4 human leukocyte antigen-A and human leukocyte antigen-B antigens tested, except B*27:05 and B*44:02. Serum absorbed with B*51:01-transfected HYM2.C1R cells left only reactivity to B17 (B57, B58), but not to any other Bw4 antigens., Conclusion: A Bw4 patient made antibody to a Bw4 epitope. This finding indicates that apparent anti-Bw4 or anti-Bw6 antibody should not be ignored even in patients who express a common Bw4 or Bw6 antigen, respectively.

Morphologic heterogeneity among melanocytic proliferations is a common challenge in the diagnosis of melanoma. In particular, atypical melanocytic lesions in children, adolescents, and young adults may be difficult to classify because of significant morphologic overlap with melanoma. Recently a four-probe fluorescence in situ hybridization (FISH) protocol to detect chromosomal abnormalities in chromosomes 6 and 11 has shown promise for improving the classification of melanocytic lesions. We sought to determine the correlation between FISH results, morphology, and clinical outcomes in a series of challenging melanocytic proliferations in young patients. We retrospectively performed the standard four-probe FISH analysis on 21 melanocytic neoplasms from 21 patients younger than 25 years of age (range 5-25 years, mean 14.6 years) from Stanford University Medical Center who were prospectively followed for a median of 51 months (range 1-136 months). The study cohort included patients with 5 confirmed melanomas, 2 melanocytic tumors of uncertain malignant potential (MelTUMPs), 10 morphologically challenging atypical Spitz tumors (ASTs), and 4 typical Spitz nevi. FISH detected chromosomal aberrations in all five melanomas and in one MelTUMP, in which the patient developed subsequent lymph node and distant metastasis. All 10 ASTs, 4 Spitz nevi, and 1 of 2 MelTUMPs were negative for significant gains or losses in chromosomes 6 and 11q. Our findings demonstrated a strong correlation between positive FISH results and the histomorphologic impression of melanoma. This finding was also true for the MelTUMP with poor clinical outcome. Therefore FISH may serve as a helpful adjunct in the classification of controversial melanocytic tumors in young patients., (© 2014 The Authors. Pediatric Dermatology published by Wiley Periodicals, Inc.)

Syngeneic graft-versus-host disease (SGVHD), a chronic inflammatory disease, develops following irradiation, syngeneic bone marrow transplantation (BMT) and treatment with the immunosuppressive agent cyclosporine A (CsA). We have shown that TH1 and TH17 cytokine responses are increased during the development of SGVHD. The current study was designed to further investigate the involvement of TH17 immunity in SGVHD-associated colitis. IL-23 is a TH17 cytokine responsible for maintaining the effector functions of TH17 cells. The administration of anti-mouse IL-23p19 was shown to significantly reduce the clinical symptoms of primary and secondary SGVHD-associated colitis resulting in a significant reduction in both TH1 and TH17 associated cytokine expression. These results demonstrate that the TH17-associated cytokine, IL-23, may prove to be a beneficial therapeutic target in the treatment of chronic colon inflammation., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a representative of a large group of polyhalogenated aromatic hydrocarbons that are widespread environmental contaminants. Administration of TCDD to laboratory animals or cultured cells results in a number of adverse effects that are well documented. For example, the effects of TCDD observed in developing organisms indicate that exposure to this class of environmental contaminants significantly alters embryo morphogenesis. However, it is not clear whether tissue regeneration in adult animals may be similarly affected. With this in mind, we examined the impact of TCDD exposure on wound healing using a murine cutaneous wound healing model. Our results indicate that TCDD exposure did not significantly alter the time needed for wound closure. However, in the TCDD-treated mice, a significant decrease in tensile strength in the healed wounds was observed which is indicative of an aberrantly healed wound. Immunostaining revealed that exposure to TCDD increased the population of macrophages detected within the wounded tissue at the latter stages of wound healing. Our findings support the idea that exposure to environmental contaminants such as TCDD is proinflammatory in the wounded tissue, disrupts normal healing and ultimately produces in a poorly healed wound., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

A case of lymphoma of T-cell derivation with aberrant expression of three B-cell lineage markers (CD19, CD20, and CD79a), which was diagnosed on a left axillary excision, is described. Immunohistochemical studies and flow cytometry analysis demonstrated neoplastic cells expressing CD3, CD19, CD20, and CD79a with absence of CD4, CD8, CD10, CD30, CD34, CD56, CD68, TDT, MPO, PAX-5, and surface immunoglobulin. Gene rearrangement studies performed on paraffin blocks demonstrated monoclonal T-cell receptor gamma chain rearrangement with no evidence of clonal heavy chain rearrangement. The neoplastic cells were negative for Epstein-Barr virus (EBV) or Human Herpes Virus 8 (HHV-8). At the time of diagnosis, the PET scan demonstrated hypermetabolic neoplastic cells involving the left axilla, bilateral internal jugular areas, mediastinum, right hilum, bilateral lungs, and spleen. However, bone marrow biopsy performed for hemolytic anemia revealed normocellular bone marrow with trilineage maturation. The patient had no evidence of immunodeficiency or infection with EBV or HHV-8. This is the first reported case of a mature T-cell lymphoma with aberrant expression of three B-cell lineage markers. The current report also highlights the need for molecular gene rearrangement studies to determine the precise lineage of ambiguous neoplastic clones.

Background: The pathogenesis of inflammatory bowel disease (IBD) is complex and the need to identify molecular biomarkers is critical. Epithelial cells play a central role in maintaining intestinal homeostasis. We previously identified five "signature" biomarkers in colonic epithelial cells (CEC) that are predictive of disease phenotype in Crohn's disease. Here we investigate the ability of CEC biomarkers to define the mechanism and severity of intestinal inflammation., Methods: We analyzed the expression of RelA, A20, pIgR, tumor necrosis factor (TNF), and macrophage inflammatory protein (MIP)-2 in CEC of mice with dextran sodium sulfate (DSS) acute colitis or T-cell-mediated chronic colitis. Factor analysis was used to combine the five biomarkers into two multifactorial principal components (PCs). PC scores for individual mice were correlated with disease severity., Results: For both colitis models, PC1 was strongly weighted toward RelA, A20, and pIgR, and PC2 was strongly weighted toward TNF and MIP-2, while the contributions of other biomarkers varied depending on the etiology of inflammation. Disease severity was correlated with elevated PC2 scores in DSS colitis and reduced PC1 scores in T-cell transfer colitis. Downregulation of pIgR was a common feature observed in both colitis models and was associated with altered cellular localization of pIgR and failure to transport IgA., Conclusions: A multifactorial analysis of epithelial gene expression may be more informative than examining single gene responses in IBD. These results provide insight into the homeostatic and proinflammatory functions of CEC in IBD pathogenesis and suggest that biomarker analysis could be useful for evaluating therapeutic options for IBD patients., (Copyright © 2012 Crohn's & Colitis Foundation of America, Inc.)

Syngeneic graft vs. host disease (SGVHD) was first described as a graft vs. host disease-like syndrome that developed in rats following syngeneic bone marrow transplantation (BMT) and cyclosporin A (CsA) treatment. SGVHD can be induced by reconstitution of lethally irradiated mice with syngeneic bone marrow cells followed by 21 days of treatment with the immunosuppressive agent CsA. Clinical symptoms of the disease appear 2-3 wk following cessation of CsA therapy, and disease-associated inflammation occurs primarily in the colon and liver. CD4(+) T cells have been shown to play an important role in the inflammatory response observed in the gut of SGVHD mice. Time-course studies revealed a significant increase in migration of CD4(+) T cells into the colon during CsA therapy, as well as significantly elevated mRNA levels of TNF-α, proinflammatory chemokines, and cell adhesion molecules in colonic tissue of CsA-treated animals compared with BMT controls, as early as day 14 post-BMT. Homing studies revealed a greater migration of labeled CD4(+) T cells into the gut of CsA-treated mice at day 21 post-BMT than control animals via CsA-induced upregulation of mucosal addressin cell adhesion molecule. This study demonstrates that, during the 21 days of immunosuppressive therapy, functional mechanisms are in place that result in increased homing of CD4(+) T effector cells to colons of CsA-treated mice.

Murine syngeneic graft-versus-host disease (SGVHD) results in chronic colon and liver inflammation following syngeneic bone marrow transplantation (BMT) and treatment with the calcineurin inhibitor, cyclosporine A (CsA). SGVHD was initially thought to arise as a result of an autoreactive immune response, but more recently it has been shown that enhanced antimicrobial responses develop in SGVHD mice. Consequently, we performed studies to analyze the role of the microbiota in the development of murine SGVHD. Treatment with broad-spectrum antibiotics eliminated disease-associated inflammatory immune responses and pathology, linking the role of the microbiota and microbial-specific immunity to the development of murine SGVHD. In a broader context, these results bring into question the role that anti-microbial immune responses play in post-transplant immune pathologies that develop following allogeneic stem cell transplantation and use of calcineurin inhibitors.

Murine syngeneic graft-versus-host disease (SGVHD) initiates colon and liver inflammation following lethal irradiation, reconstitution with syngeneic bone marrow transplantation, and therapy with the immunosuppressive agent cyclosporine A. Previous studies have demonstrated that the inducible disease is mediated by CD4(+) T cells with increased reactivity of peripheral and liver-associated lymphocytes against intestinal microbial Ags. In the current report, studies were performed to analyze the specificity of the CD4(+) T cell response of T cells isolated from diseased animals and to determine the in vivo role of the microbiota to the development of SGVHD. Increased major histocompatibility Ag (MHC) class II-restricted responsiveness of SGVHD CD4(+) T cells against microbial Ags isolated from the ceca of normal animals was observed. The enhanced proliferative response was observed in the CD62L(-) memory population of CD4(+) T cells. To determine the role of the bacterial microbiota in the development of murine SGVHD, control and CsA-treated bone marrow transplantation animals were treated with broad-spectrum antibiotics (metronidazole, ciprofloxacin) after transplantation. Cyclosporine A-treated animals that were given antibiotic therapy failed to develop clinical symptoms and pathological lesions in the target tissues characteristic of SGVHD. Furthermore, the reduction in intestinal bacteria resulted in the elimination of the enhanced antimicrobial CD4(+) T cell response and significantly reduced levels of the inflammatory cytokines, IFN-γ, IL-17, and TNF-α. The elimination of the disease-associated inflammatory immune responses and pathology by treatment with broad-spectrum antibiotics definitively links the role of the microbiota and microbial-specific immunity to the development of murine SGVHD.

Although Congress recently passed health insurance reform legislation, the real catalyst for change in the health care delivery system, the author's argue, will be changes to the reimbursement model. To rein in increasing costs, the Centers for Medicare and Medicaid aims to move Medicare from the current fee-for-service model to a reimbursement approach that shifts the risk to providers and encourages greater accountability both for the cost and the quality of care. This level of increased accountability can only be achieved by clinical integration among health care providers. Central to this reorganized delivery model are primary care providers who coordinate and organize the care of their patients, using best practices and evidence-based medicine while respecting the patient's values, wishes, and dictates. Thus, the authors ask whether primary care physicians will be available in sufficient numbers and if they will be adequately and appropriately trained to take on this role. Most workforce researchers report inadequate numbers of primary care doctors today, a shortage that will only be exacerbated in the future. Even more ominously, the authors argue that primary care physicians being trained today will not have the requisite skills to fulfill their contemplated responsibilities because of a variety of factors that encourage fragmentation of care. If this training issue is not debated vigorously to determine new and appropriate training approaches, the future workforce may eventually have the appropriate number of physicians but inadequately trained individuals, a situation that would doom any effort at system reform.

Syngeneic graft-versus-host disease (SGVHD) develops following lethal irradiation, reconstitution with syngeneic bone marrow (BM) and treatment with a 21 day course of the immunosuppressive agent cyclosporine A (CsA). Clinical symptoms of SGVHD appear 2-3 weeks post-CsA with inflammation occurring in the colon and liver. Previously we have demonstrated that CD4(+) T cells and a T helper cell type 1 cytokine response (T(H)1) are involved in the development of SGVHD associated intestinal inflammation. Studies have recently discovered an additional T cell lineage that produces IL-17 and is termed T(H)17. It has been suggested that inflammatory bowel disease is a result of a T(H)17 response rather than a T(H)1 response. This study was designed to investigate T(H)17 involvement in SGVHD-associated colitis. Following induction of SGVHD, the levels of T(H)17 and T(H)1 cytokine mRNA and protein were measured in control and SGVHD animals. In vivo cytokine neutralization was performed to determine the role of the prototypic T(H)17 cytokine, IL-17, in the disease process. We found that during CsA-induced murine SGVHD there was an increase in both T(H)17 and T(H)1 mRNA and cytokines within the colons of diseased mice. The administration of an anti-mouse IL-17A mAb did not alter the course of disease. However, neutralization of IL-17A resulted in an increased production of IL-17F, a related family member, with an overlapping range of effector activities. These results demonstrate that in the pathophysiology of SGVHD, there is a redundancy in the T(H)17 effector molecules that mediate the development of SGVHD., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The murine model of cyclosporine A (CsA)-induced syngeneic graft-versus-host disease (SGVHD) is a bone marrow (BM) transplantation model that develops chronic colon inflammation identical to other murine models of CD4(+) T cell-mediated colitis. Interestingly, SGVHD animals develop chronic liver lesions that are similar to the early peribiliary inflammatory stages of clinical chronic liver disease, which is frequently associated with inflammatory bowel disease (IBD). Therefore, studies were initiated to investigate the chronic liver inflammation that develops in the SGVHD model. To induce SGVHD, mice were lethally irradiated, reconstituted with syngeneic BM, and treated with CsA. All of the SGVHD animals that developed colitis also develop chronic liver inflammation. Liver samples from control and SGVHD animals were monitored for tissue pathology, RNA for inflammatory mediators, and phenotypic analysis and in vitro reactivity of the inflammatory infiltrate. Diseased animals developed lesions of intrahepatic and extrahepatic bile ducts. Elevated levels of mRNA for molecules associated with chronic liver inflammation, including mucosal cellular adhesion molecule -1, the chemokines CCL25, CCL28, CCR9, and T(H)1- and T(H)17-associated cytokines were observed in livers of SGVHD mice. CD4(+) T cells were localized to the peribiliary region of the livers of diseased animals, and an enhanced proliferative response of liver-associated mononuclear cells against colonic bacterial antigens was observed. The murine model of SGVHD colitis may be a valuable tool to study the entero-hepatic linkage between chronic colon inflammation and inflammatory liver disease.

Background: Previously we have shown that B cell receptor (BCR) expression and B cell receptor signaling pathways are important for the basal growth of B lymphoma cells. In particular we have shown that the activation of Syk, a non-src family protein tyrosine kinase and the mitogen activated protein kinases (MAPK), ERK and JNK that mediate BCR signals are required for the constitutive growth of B lymphoma cells. Since src family protein tyrosine kinases (SFKs) like Lyn are known to be needed for the phosphorylation of BCR co-receptors, Ig-alpha and Ig-beta, we hypothesized that one or more SFKs will be constitutively activated in B lymphoma cells and may be necessary for B lymphoma growth., Results: Src kinase activity was found to be constitutively high in many murine and human B lymphoma cell lines and primary lymphoma samples. The specific pharmacological inhibitors of SFKs, PP1 and PP2 inhibited the proliferation of a number of both murine and human B lymphomas in a dose-dependent manner. Importantly, dasatinib (BMS-354825), an oral dual BCR-ABL and SFK specific inhibitor inhibited the growth of B lymphomas in the nanomolar range in vitro and strongly inhibited a mouse lymphoma growth in vivo. Among the SFKs, Lyn is predominantly phosphorylated and Lyn-specific small interfering RNA inhibited the growth of B lymphomas, supporting an important role for Lyn in B lymphoma growth. Suppression of SFK activity blocks BCR mediated signaling pathways. PMA or CpG can partially reverse the growth inhibition induced by SFK inhibition. Although blocking SFK activity inhibited the growth of a number of B lymphomas, some lymphomas such as SudHL-4, SudHL-6, OCI-Ly3 and OCI-Ly10 are more resistant due to an increased expression of the anti-apoptotic proteins Bcl-2 and Bcl-xL., Conclusions: These studies further support our concept that BCR signaling pathways are important for the continued growth of established B lymphoma cells. Some of the intermediates in this BCR pathway are potential immunotherapeutic targets. In particular, inhibition of SFK activity alone or in synergy with inhibition of the prosurvival Bcl-2 proteins holds promise in developing more effective treatments for B lymphoma patients.

Purpose: To evaluate the subjective and objective results of surface replacement arthroplasty (SRA) for arthritis of the proximal interphalangeal (PIP) joint using the PIP-SRA implant. Emphasis is placed on causes of complications, failures, and techniques used for revision., Methods: This is a retrospective review of 43 surface replacement PIP joint arthroplasties performed in 25 patients using the PIP-SRA implant. Subjective results were obtained through a mailed questionnaire. Pre- and postoperative ranges of motion were obtained for PIP joints and DIP joints. X-rays were evaluated for signs of subsidence, periprosthetic radiolucency, loosening, or stress-shielding. Joints requiring revision were separately analyzed., Results: The average follow-up time was 37 months (range, 12 to 72 months). The average active PIP joint arc of motion went from 57 degrees before surgery to 58 degrees after surgery, excluding 2 joints that were salvaged with arthrodesis. The average active DIP joint arc of motion went from 36 degrees before surgery to 24 degrees after surgery, excluding arthrodeses. Satisfaction rating revealed 26 very satisfactory (60%), 12 fairly satisfactory (28%), and 5 not satisfactory (12%). Thirty-three patients rated their joint pain better, 3 joints were unchanged, and 7 were worse. Eleven (26%) arthroplasties failed, requiring major revision (arthrodesis or replacement of 1 or both components) for pain. Ten of 11 revisions were due to loosening associated with the lack of cement. Revision procedures produced satisfactory results in 8 of 11 joints., Conclusions: Surface replacement arthroplasty of the PIP joint holds promise for the future. It offers motion and stability for the index finger unattainable with silicone arthroplasty. Our results do not differ notably from those of other series using this implant, except that failures due to loosening in our study were almost exclusively associated with the lack of cement. Therefore, we recommend using cement with the PIP-SRA implant in every case until superior long-term results can be demonstrated using uncemented components. Proximal interphalangeal joint arthroplasty is an exacting procedure no matter what technique or implant is used, and no one technique has yet been proven superior to all others., Type of Study/level of Evidence: Therapeutic IV.

Purpose: Responding to suggestions that physicians are obligated to inquire fully about complementary and alternative medicine (CAM) use and its scientific evidence, to acknowledge patients' health beliefs and practices, and to accommodate diverse healing practices, our interdisciplinary CAM integration project created an advisory committee (AC) composed of CAM practitioners and institutional personnel to incorporate CAM- related information into health professions training. We report on the collaborative process and describe group members' perceptions of medicine and clinical teaching., Methods: Information collected from the first two years' quarterly meetings, the first annual retreat, and other venues was analyzed in conjunction with semi-structured in-person interviews of 10 biomedical and CAM practitioner committee members. Data were analyzed using qualitative methodology and N5 software to identify themes and patterns., Results: Analysis confirmed expectations that allopathic and CAM AC members held different views of health and healing. Member comments reflected points of tension that clustered into three intertwined themes: what constitutes evidence , interaction with the patient , and the relative importance of experience in learning . Recommendations for designing interdisciplinary CAM curricula are presented., Conclusion: Differences between CAM and allopathic providers were frequent but did not obviate common goals or collaboration. Results demonstrate the potential for collaboration between these groups and our activities may be useful to others seeking to implement interdisciplinary care, particularly between CAM and allopathic providers.

Acute myeloid leukemia (AML) is an aggressive hematological cancer. Despite therapeutic regimens that lead to complete remission, the vast majority of patients undergo relapse. The molecular mechanisms underlying AML development and relapse remain incompletely defined. To explore whether loss of DNA mismatch repair (MMR) function is involved in AML, we screened two key MMR genes, MSH2 and MLH1, for mutations and promoter hypermethylation in leukemia specimens from 53 AML patients and blood from 17 non-cancer controls. We show here that whereas no amino acid alteration or promoter hypermethylation was detected in all control samples, 18 AML patients exhibited either mutations in MMR genes or hypermethylation in the MLH1 promoter. In vitro functional MMR analysis revealed that almost all the mutations analyzed resulted in loss of MMR function. MMR defects were significantly more frequent in patients with refractory or relapsed AML compared with newly diagnosed patients. These observations suggest for the first time that the loss of MMR function is associated with refractory and relapsed AML and may contribute to disease pathogenesis.

Syngeneic graft-vs.-host disease (SGVHD) develops in rodents following the treatment of lethally irradiated, bone marrow (BM) reconstituted animals with a short course of the immunosuppressive agent cyclosporine A (CsA). Using an in vivo depletion approach, we recently demonstrated that CD4(+), but not CD8(+), T cells participated in inducing SGVHD. Studies were therefore undertaken to adoptively transfer SGVHD into lethally irradiated, syngeneic BM reconstituted secondary recipients. Whole T cell populations as well as purified CD4(+)T cells isolated from SGVHD, but not normal or transplant control, animals mediated the transfer of SGVHD into secondary recipients. These cells have an apparent specificity for enteric bacterial antigens. The pathologic process that developed was identical to that observed in the animals with de novo SGVHD after syngeneic BMT and CsA therapy. It was shown that a radiation-sensitive mechanism prevented the transfer of SGVHD into normal, nonirradiated secondary recipients. The ability to reproducibly transfer SGVHD into secondary recipients will enhance our ability to study regulatory mechanisms that are altered during CsA therapy and permit the development of murine CsA-induced SGVHD.

Prostate apoptosis response-4 (Par-4) is a tumor-suppressor protein that induces apoptosis in cancer cells, but not in normal/immortalized cells. The cancer-specific proapoptotic action of Par-4 is encoded in its centrally located SAC domain. We report here the characterization of a novel mouse model with ubiquitous expression of the SAC domain. Although SAC transgenic mice displayed normal development and life span, they were resistant to the growth of spontaneous, as well as oncogene-induced, autochthonous tumors. Resistance to tumorigenesis was linked to inhibition of nuclear factor-kappaB activity and induction of apoptosis by the SAC domain. Collectively, our findings provide genetic evidence that the SAC domain of Par-4 confers cancer resistance in transgenic mice without compromising normal viability or aging, and may have therapeutic significance.

Background: Syngeneic graft-versus-host disease (SGVHD) develops after lethal irradiation, reconstitution with syngeneic bone marrow (BM), and treatment with a 21-day course of the immunosuppressant cyclosporine A (CsA). Clinical symptoms of SGVHD appear 2-3 weeks after CsA treatment, with inflammation in the colon and liver. It has been demonstrated that CD4+ T cells and a T helper cell type 1 cytokine response (Th1) are involved in the development of SGVHD associated intestinal inflammation. The immune response associated with SGVHD is thought to be the result of the reconstitution of the recipient immune system with the syngeneic donor BM. However, definitive studies have not addressed this issue experimentally., Methods: To determine the origin of the effector cells that participate in SGVHD, C3H/HeN recipient mice were lethally irradiated and transplanted with BM from normal immunocompetent mice or from immunodeficient, severe combined immune deficient, or Rag-2-/- animals., Results: CsA-treated animals, but not control animals, developed inflammation characteristic of SGVHD in the colon and liver regardless of the source of the donor marrow. Furthermore, immunologically, all CsA treated animals responded similarly with increased production of inflammatory cytokines and an increase in activated CD4+ T cells in the periphery and colon relative to controls., Conclusion: These results demonstrate that after lethal irradiation and in the absence of donor T cells, T cells of recipient origin can expand and mediate the induction of CsA-induced SGVHD.

As medical education grapples with organizational calls for centralized curricular oversight, programs may be compelled to respond by establishing highly vertical, stacked governance structures. Although these models offer discrete advantages over the horizontal, compartmentalized structures they are designed to replace, they pose new challenges to ensuring curricular quality and the educational innovations that drive the curricula. The authors describe a hybrid quality-assurance (QA) governance structure introduced in 2003 at the University of Kentucky College of Medicine (UKCOM) that ensures centralized curricular oversight of the educational product while allowing individualized creative control over the educational process. Based on a Lean production model, this approach draws on industry experiences that strategically separate institutional accountability (management) for a quality curriculum from the decision-making processes required to ensure it (production). In so doing, the authors acknowledge general similarities and key differences between overseeing the manufacture of a complex product versus the education of a physician-emphasizing the structured, sequential, and measurable nature of each process. Further, the authors briefly trace the emergence of quality approaches in manufacturing and discuss the philosophical changes that accompany transition to an institutional governance system that relies on vigorous, robust performance measures to offer continuous feedback on curricular quality.

Curcumin (diferuloylmethane), a component of dietary spice turmeric (Curcuma longa), has been shown in recent studies to have therapeutic potential in the treatment of cancer, diabetes, arthritis, and osteoporosis. We investigated the ability of curcumin to modulate the growth of B lymphomas. Curcumin inhibited the growth of both murine and human B lymphoma in vitro and murine B lymphoma in vivo. We also demonstrate that curcumin-mediated growth inhibition of B lymphoma is through inhibition of the survival kinase Akt and its key target Bad. However, in vitro kinase assays show that Akt is not a direct target of curcumin. We identified a novel target for curcumin in B lymphoma viz spleen tyrosine kinase (Syk). Syk is constitutively activated in primary tumors and B lymphoma cell lines and curcumin down-modulates Syk activity accompanied by down-regulation of Akt activation. Moreover, we show that overexpression of Akt, a target of Syk, or Bcl-x(L), a target of Akt can overcome curcumin-induced apoptosis of B lymphoma cells. These observations suggest a novel growth promoting role for Syk in lymphoma cells.

B lymphomas account for the majority of the lymphoma cases. BCR expression appears to be important for B lymphoma because most oncogenes are translocated to nonrearranged Ig loci and because all of the variants that arise in anti-idiotypic Ab-treated lymphoma patients remain BCR positive. Based on this and the fact that BCR is required for mature B cell survival, we tested the requirement for continued expression of BCR for the growth and survival of B lymphoma cells. Using Igalpha or Igbeta-specific small interfering RNA (siRNA) to inhibit BCR expression, we demonstrate for the first time that constitutive signaling by BCR is critical for survival and proliferation of both murine and human B lymphoma cells. The BCR signals in lymphoma appear to be mediated by Syk, as it is constitutively active in a variety of B lymphoma cells. Blocking Syk activity by selective inhibitors suppresses growth of several murine and human B lymphomas.

The University of Kentucky College of Medicine (UKCOM) retains a long history of educational commitment, quality, and innovation. Since undergoing a major curricular revision in the early 1990s, the evolving UKCOM curriculum has continued to incorporate advances in biomedical knowledge and pedagogy while meeting changing societal needs and expectations for physicians in practice. Building upon its established record of excellence in medical education, a curricular quality assurance (QA) program has been initiated to more efficiently guide improvement and innovation by providing faculty with key resources to identify and disseminate local best practices in teaching, learning, and evaluation. Through such efforts, the University of Kentucky College of Medicine is committed to providing a rich educational experience that will meet the needs of students, physicians, and patients well into the 21st century.

Several primary murine and human B lymphomas and cell lines were found to constitutively express high levels of the activated form of c-jun N-terminal kinase (JNK), a member of the mitogen-activated protein (MAP) kinase family. Proliferation of murine B lymphomas CH31, CH12.Lx, BKS-2, and WEHI-231 and the human B lymphomas BJAB, RAMOS, RAJI, OCI-Ly7, and OCI-Ly10 was strongly inhibited by SP600125, an anthrapyrazolone inhibitor of JNK, in a dose-dependent manner. The lymphoma cells underwent apoptosis and arrested at the G2/M phase of cell cycle. Furthermore, JNK-specific small interfering RNA (siRNA) inhibited the growth of both murine and human B lymphomas. Thus in the B-lymphoma model, JNK appears to have a unique prosurvival role. Survival signals provided by CD40 and interleukin-10 (IL-10) together reversed the growth inhibition induced by the JNK inhibitor. c-Myc protein levels were reduced in the presence of both SP600125 and JNK-specific siRNA, and CD40 ligation restored c-Myc levels. Moreover, Bcl-xL rescued WEHI-231 cells from apoptosis induced by the JNK inhibitor. The JNK inhibitor also reduced levels of early growth response gene-1 (Egr-1) protein, and overexpressing Egr-1 partially rescued lymphoma cells from apoptosis. Thus, JNK may act via c-Myc and Egr-1, which were shown to be important for B-lymphoma survival and growth.

Syngeneic graft-vs-host disease (SGVHD) develops following lethal irradiation, reconstitution with syngeneic bone marrow, and treatment with a 21-day course of the immunosuppressive agent cyclosporin A (CsA). Following cessation of CsA, this inducible disease is characterized by weight loss, diarrhea, and development of inflammation in the colon and liver. Although nonspecific effector cells and Th1 cytokines have been shown to participate in disease induction, the role of T cells has not been fully elucidated. Initial studies demonstrated significant increases in CD4+ T cells, but not other T cell populations in the colons of diseased animals relative to transplant control animals. To demonstrate a functional linkage between increases in colonic CD4+ T cells and disease induction, in vivo T cell depletion studies were performed. Beginning on the day of bone marrow transplantation, groups of control and CsA-treated animals were treated with mAb against either CD4 or CD8 for 21 days. Treatment with anti-CD4, but not anti-CD8, eliminated clinical symptoms and colon pathology. Interestingly, neither anti-CD4 nor anti-CD8 therapy affected the development of liver pathology associated with SGVHD. These findings demonstrated that CD4+ T cells initiate development of the intestinal inflammation associated with murine SGVHD.

Oxidative stress enhanced by transition metals such as iron forms an attractive hypothesis for neurodegeneration in Alzheimer's Disease (AD). Iron is increased in the brain in AD, but whether this is a primary abnormality or the result of secondary accumulation is unclear. Among several genetic loci associated with AD, the locus at chromosome 6p21 contains the hereditary hemochromatosis gene HFE. To determine whether a genetic predisposition to iron accumulation is associated with AD, we evaluated three hemochromatosis-associated HFE mutations and APOE in cognitively and histopathologically evaluated subjects with AD, mild cognitive impairment (MCI), non-demented controls with AD-like pathologic changes defined by Braak stage > or = 3 (high pathology controls (HPC)), and non-demented controls without significant histologic changes (low-pathology controls (LPC)). In a subset, we examined ventricular (CSF) fluid F(2)-isoprostane (F(2)-IsoP) levels, a marker of lipid peroxidation. Seventeen subjects demonstrated homozygous or compound heterozygous HFE mutations, 13 (9.4%) in the AD/MCI group (P = 0.019 vs. LPC) and four (20%) in the HPC group (P = 0.006, P < 0.05 with Bonferroni correction vs. LPC). In contrast, the APOE4 allele frequency was increased only in the AD/MCI patients (P < 10(-3) vs. HPC, P < 10(-6) vs. LPC). F(2)-IsoP levels were increased in AD subjects with any HFE mutation versus wild type HFE (P = 0.027). Although confirmation is required, these findings suggest that HFE mutations are associated with increased oxidative stress and Braak stage, and that HFE and APOE genotypes are different between AD patients, high pathology and low pathology controls., (Copyright 2003 Wiley-Liss, Inc.)

Syngeneic graft-versus-host disease (SGVHD) develops following lethal irradiation, reconstitution with syngeneic bone marrow, and treatment with a short course of cyclosporin A (CsA) therapy. The disease is characterized by the development of a T helper cell type 1-like cytokine response [interleukin (IL)-12, interferon-gamma (IFN-gamma), and tumor necrosis factor alpha], and macrophage activation is central to development of the syndrome. It has been shown that nitric oxide (NO) participates significantly in the development of allogeneic GVHD. Studies were initiated to determine if NO participates in the pathology associated with SGVHD. Significant increases in inducible NO synthase (iNOS) mRNA and circulating NO were found in the tissues of SGVHD versus control animals. Treatment of SGVHD animals with the iNOS inhibitor aminoguanidine (AG) reversed the pathology associated with this disease. Furthermore, AG treatment reduced the production of IL-12 and IFN-gamma mRNA in the colons of CsA-treated mice. These studies demonstrate that NO participates in the pathological processes that are associated with the development of murine SGVHD.

Folate is a critical factor for DNA metabolism and its deficiency is associated with a number of human diseases and cancers. Although it has been shown that folate deficiency induces genomic instability and apoptotic cell death, the underlying mechanism is largely unknown. Given the role of mismatch repair in maintaining genomic integrity, mismatch repair was tested for its involvement in folate deficiency-induced genomic instability and cell death. Cells proficient in mismatch repair were highly sensitive to folate deficiency compared with cells defective in either hMutSalpha or hMutLalpha. Since wild-type cells but not mutant cells underwent apoptosis upon extensive folate depletion, the apoptotic response is dependent on a functional mismatch repair system. Our data also indicate that p53 is required for the folate depletion-induced apoptosis. In vitro biochemical studies demonstrated that hMutSalpha specifically recognized DNA damage induced by folate deficiency, suggesting a direct participation of mismatch repair proteins in mediating the apoptotic response. We conclude that while the mismatch repair-dependent apoptosis is necessary to protect damaged cells from tumorigenesis, it may damage a whole tissue or organ, as seen in patients with megaloblastic anemia, during extensive folate deficiency.

Two different doses of Ross River virus (RR) were fed to Ochlerotatus vigilax (Skuse), the primary coastal vector in Australia; and blood engorged females were held at different temperatures up to 35 d. After ingesting 10(4.3) CCID50/mosquito, mosquitoes reared at 18 and 25 degrees C (and held at the same temperature) had higher body remnant and head and salivary gland titers than those held at 32 degrees C. although infection rates were comparable. At 18, 25, and 32 degrees C, respectively, virus was first detected in the salivary glands on days 3, 2, and 3. Based on a previously demonstrated 98.7% concordance between salivary gland infection and transmission, the extrinsic incubation periods were estimated as 5, 4, and 3 d, respectively, for these three temperatures. When Oc. vigilax reared at 18, 25, or 32 degrees C were fed a lower dosage of 10(3.3) CCID50 RR/mosquito, and assayed after 7 d extrinsic incubation at these (or combinations of these) temperatures, infection rates and titers were similar. However, by 14 d, infection rates and titers of those reared and held at 18 and 32 degrees C were significantly higher and lower, respectively. However, this process was reversible when the moderate 25 degrees C was involved, and intermediate infection rates and titers resulted. These data indicate that for the strains of RR and Oc. vigilax used, rearing temperature is unimportant to vector competence in the field, and that ambient temperature variations will modulate or enhance detectable infection rates only after 7 d extrinsic incubation. Because of the short duration of extrinsic incubation, however, this will do little to influence RR epidemiology, because by this time some Oc. vigilax could be seeking their third blood meal, the latter two being infectious.

Syngeneic GVHD (SGVHD) develops following syngeneic bone marrow transplantation and treatment with cyclosporine A. Previous studies have demonstrated a role for IL-12, IFN-gamma, and TNF-alpha in the development of murine SGVHD. Macrophages can be activated to secrete IL-12 and TNF-alpha via a T-cell-dependent or T-cell-independent pathway (LPS or bacterial products). Studies were designed to determine if LPS participated in the development of SGVHD in C3H/HeN (LPS-responsive) and C3H/HeJ (LPS-hyporesponsive) mice. C3H/HeJ and C3H/HeN mice had similar levels of disease induction and pathology. Following induction of SGVHD, treatment of C3H/HeN, but not C3H/HeJ, mice with a sublethal dose of LPS resulted in mortality. However, neutralization of IL-12 abrogated the development of disease in C3H/HeJ mice, demonstrating that activated macrophages and their products participated in the development of SGVHD in these animals. These data suggested that LPS responsiveness was not a predisposing factor for SGVHD induction.

This brief communication describes the characterization of a new allele, DRB1*1336.